Compositions and methods for rendering tumor cells susceptible to CD8+ T cell-mediated killing

Abstract

The present invention provides an immunoconjugate having the formula: T-c-E n -c-Fc n or T-c-Fc n -c-E n ; wherein, T is a single chain variable portion fragment of a monoclonal antibody (scFv) directed to a target protein, polypeptide, or fragment thereof, which is highly expressed on cancer cells; E is two or more foreign immunogenic CD8 + T cell antigenic epitopes; c is a peptide or polypeptide fragment thereof, capable of being cleaved by a specific protease; and Fc is two or more Fc portions of an IgG antibody. Nucleic acid sequences encoding the same and vectors containing said nucleic acid sequences are also provided. Methods of making the immunoconjugate, along with methods of making target cells susceptible to CTL mediated cell killing, and methods for treatment of cancers are also provided.

The invention claimed is: 
     
       1. A method for making a tumor cell susceptible to CD8 +  T cell killing, comprising contacting one or more tumor cells with an effective amount of an immunoconjugate having the formula:
   T-c-E n -c-Fc n ; 
 wherein 
 T is a single chain variable portion fragment of a monoclonal antibody (scFv) directed to a target protein, polypeptide, or fragment thereof, which is highly expressed on cancer cells, and selected from the group consisting of scFv directed to mesothelin, epidermal growth factor receptor (EGFR), NKG2D, or Her2/neu protein; 
 E n  is two or more foreign immunogenic CD8 +  T cell antigenic epitopes derived from ovalbumin, Epstein-Barr virus, cytomegalovirus, human papilloma virus, and influenza wherein n is 1 to 10; 
 c is a peptide or polypeptide fragment thereof, capable of being cleaved by by furin, MMP1 or MMP9; and 
 Fc n  is two or more Fc portions of an IgG antibody wherein n is 1 to 10. 
 
     
     
       2. A method for making a tumor cell susceptible to CD8 +  T cell killing, comprising contacting one or more tumor cells with an effective amount of an immunoconjugate having the formula:
   T-c-Fc n -c-E n ; 
 wherein 
 T is a single chain variable portion fragment of a monoclonal antibody (scFv) directed to a target protein, polypeptide, or fragment thereof, which is highly expressed on cancer cells, and selected from the group consisting of scFv directed to mesothelin, epidermal growth factor receptor (EGFR), NKG2D, or Her2/neu protein; 
 E n  is two or more foreign immunogenic CD8 +  T cell antigenic epitopes derived from ovalbumin, Epstein-Barr virus, cytomegalovirus, human papilloma virus, and influenza wherein n is 1 to 10; 
 c is a peptide or polypeptide fragment thereof, capable of being cleaved by by furin, MMP1 or MMP9; and 
 Fc n  is two or more Fc portions of an IgG antibody wherein n is 1 to 10. 
 
     
     
       3. The method of  claim 1 , wherein c is a furin cleavable peptide having the amino acid sequence RVKR (SEQ ID NO: 2). 
     
     
       4. The method of  claim 2 , wherein c is a furin cleavable peptide having the amino acid sequence RVKR (SEQ ID NO: 2). 
     
     
       5. The method of  claim 1 , wherein E is an ovalbumin epitope having the amino acid sequence SIINFEKL (SEQ ID NO: 1). 
     
     
       6. The method of  claim 2 , wherein E is an ovalbumin epitope having the amino acid sequence SIINFEKL (SEQ ID NO: 1). 
     
     
       7. The method of  claim 1 , wherein T is a scFv directed to mesothelin. 
     
     
       8. The method of  claim 2 , wherein T is a scFv directed to mesothelin. 
     
     
       9. The method of  claim 1 , wherein the method further comprises, determining whether the CD8 +  T cell antigenic epitopes is specific for an antigen presented on the tumor cell and then contacting one or more tumor cells with the immunoconjugate of  claim 1  having said antigenic epitope. 
     
     
       10. The method of  claim 2 , wherein the method further comprises, determining whether the CD8 +  T cell antigenic epitopes is specific for an antigen presented on the tumor cell and then contacting one or more tumor cells with the immunoconjugate of  claim 2  having said antigenic epitope. 
     
     
       11. The method of  claim 1 , wherein the tumor cell is a cancer cell. 
     
     
       12. The method of  claim 2 , wherein the tumor cell is a cancer cell. 
     
     
       13. The method of  claim 11 , wherein the tumor cell is in a subject. 
     
     
       14. The method of  claim 12 , wherein the tumor cell is in a subject.